 2011 RELS ISSN: 0974-4908

http://rels.110mb.com Res. Environ. Life Sci.

[email protected] 4(3) 133-136 (2011)

Production of bioplastic using Spirulina platensis

and comparison with commercial plastic
N. Uma Maheswari* and K. Ahilandeswari
P.G & Research Department of Microbiology, Sengamala Thayaar Educational Trust,
Women’s College, Mannargudi – 614 001, India
*e-mail: [email protected]

(Received: April 28, 2011; Revised received: August 25, 2011; Accepted: August 27, 2011)
Abstract: A lack of degradability and the closing of landfill sites, as well as growing water and land pollution problems, have led to concern about plastics.
Bioplastic was prepared by using Spirulina platensis cultivated in modified Zarrouk medium. The medium was optimized by using sodium acetate in aim
of increase the PHB concentration of S. platensis. The S. platensis was measured for total dry weight and PHB content by UV spectrophotometer in before
and after optimization. When compared the result of measurements optimized medium contain S. platensis had little higher PHB (6.20 %) content than
untreated one (5.18 %). Then S. platensis was harvested and bioplastic was prepared. In addition, chemical based commercial plastic was prepared and
compared with bioplastic for plasticizing, moldable property. Biodegrading property and cost were compared by studying the composition. The result
showed that the plasticizing and moldable property was good in all plastics. In biodegradation S. platensis bioplastic was considered as better due to
presence of PHB content but the cost of S. platensis was higher than other chemical and organic polymers.

Key words: Bioplastic, Spirulina platensis, Zarrouk medium, UV Spectrophotometer, PHB

Introduction Thermoplastics, can be melted and formed again and again;

Bioplastics or organic plastics are a form of plastics derived thermosetting plastics, once formed, are destroyed rather than melted
from renewable biomass sources such as vegetable oil, corn starch, by heating. These traditional plastics are not biodegradable.
pea starch unlike fossil-fuel plastics derived from petroleum.
Algal bioplastics are referred as plastics produced from
Biodegradable form of plastic was first characterized in the mid-
algal biomass or by using algal materials. The algal bioplastic are
1920s by French researchers. The molecule is called
easily biodegradable than compared to commercial plastics. The
polyhydroxybutyrate (PHB). Many different types of bacteria and
direct algal biomass usage for algal bioplastics is responsible for
algae produce it as food storage material (Falcone, 2004).
biodegradation. Algae based plastics have been a recent trend in
Biodegradable plastics can decompose into carbon dioxide,
the era of bioplastics (Greg stevens, 2010). The various bioplastics
methane, water, inorganic compounds, or biomass via microbial
that can be made from algae feedstock include Hybrid plastics,
assimilation. Algae serve as an excellent feedstock for plastic
Cellulose-based plastics, Poly-Lactic Acid (PLA), Bio-Polyethylene.
production owing to its many advantages such as high yield and the
Hybrid plastics are made by adding denatured algae biomass to
ability to grow in a range of environments. (Greg stevens, 2010).
petroleum based plastics like polyurethane and polyethylene as
Today, the production of plastics amounts to about 150 million fillers. The PHB content of the algal biomass increase biodegradable
tons per year and displays an upward trend (Okada, 2002). The property of plastic. It thus decreases the amount of petroleum used
main problem is environmental pollution resulting from the disposal per unit of plastic.
of polymeric materials, which may take many years to decompose.
Recycling alone has proved to be insufficient to solve this problem, In Cyanobacteria PHB was first detected in Chlorogloea
since it is impossible to recover all discarded plastic materials by this fritschii where in the presence of a reduced carbon source, it
process. A naturally produced, biodegradable form of plastic was constitutes up to 10% of dry weight. The polymer has been identified
first characterized in the mid-1920s by French researchers. The in other cyanobacteria Aphanocapsa and Spirulina spp. Spirulina
molecule is called polyhydroxybutyrate, more commonly referred spp are important PHB accumulating microorganisms. Spirulina
to as PHB. Many different types of bacteria and algae produce it as spp can be grown in variety of chemical medium, modified organic
food storage material (Falcone, 2004). medium and human excretion also used as supplement for Spirulina
growth. Spirulina platensis is one of important spp of Spirulina
The term “plastics” encompasses organic materials, such compare than other spp due to natural availability and fast growing
as the elements carbon (C), hydrogen (H), nitrogen (N), chlorine property. It can be grown in wide variety of synthetic medium. The
(Cl) and sulfur (S). The commercial plastic fall into two classes: intracellular PHB accumulating property of Spirulina platensis is

Research in Environment and Life Sciences 133 August, 2011

N. Uma Maheswari and K. Ahilandeswari Production of bioplastic

high compare then other Spirulina spp and some cyanobacteria Intracellular accumulation of PHB seems to be related to
(Costa, 2004). the ability of cyanobacteria to grow in the presence of acetate.
Normally the PHB production of organism was induced by á-
PHB was first identified by Lemoigne in 1926 as a reserve ketothiolase enzyme. The enzyme was inhibited by free coenzyme
material of Bacillus megaterium. PHB is a very common and A. However, in presence of high amount of acetyl CoA the inhibitory
widespread storage material in many micro-organisms. PHB is an effect of coenzyme A is overcome. The addition of sodium acetate to
environmentally degradable material. PHB belongs to a family of medium will monitor the acetyl CoA activity. Hence the medium was
polyesters called polyhydroxyalkanoates or PHAs. The one striking optimized by using sodium acetate. Sodium acetate was added, up
difference between PHB and petroleum-based plastics is that PHB to a final concentrations of 1g per liter. After optimization of the medium
are biodegradable. When these bioplastics are disposed to the the Spirulina platensis was allowed for determination of PHB content
environments populated by organisms such as bacteria, fungi and by UV spectrophotometry (Massimo Vincenzini et al, 1990).
algae, PHB are broken down to their essence-carbon dioxide and
water-and recycled by the natural metabolic processes of these When the cyanobacterium was floated on surface it is
microbes (Lemoigne, 1927). harvested and dried. Here the S. platensis act as a biopolymer. In
commercial plastic preparation ethylene or propylene to be converted
This present study was aimed at the production of bioplastic into a powdered polymer material. Here S. platensis converted into
and commercial plastic by using glycerol as petrol based ingredients, powdered polymer material and used as 33% concentration with
by using biopolymer and commercial polymer. Here the commercial polymer. The Spirulina bioplastic ingredients include,
cyanobacteria, Spirulina platensis used as biopolymer. The bioplastic Sorbitol 2.25g, Gelatin 2.25g, S. platensis 2.25g, Glycerol solution
and commercial plastic was compared for various properties include 180ml of 1% , Water 24 ml. All the ingredients mixed well and heat
plasticizing capacity, moldable property, biodegradable property the mixture to 95ºC. Stirred the mixture while were heating it, and
and cost requirement. once it is at the right temperature, removed the heat and keep
stirring. Poured the mixture into a dried pan, and spread it out to let
Materials and Methods
it dry. The time required for separation of plastic is depend on
Cyanobacterium, Spirulina platensis sample were collected temperature of the room, it may take several days. After complete
from Aurovil, Health centre, Pondicherry and cultivated in Zarrouk drying the bioplastic separated from the pan (Greg Stevens, 2010).
medium (Pandey, 2010) at small and large-scale level. In small
scale cultivation Spirulina was inoculated into conical flask with The commercial petrol based plastic was prepared in aim
Zarrouk medium and maintained for 10 days. During the period, of compared with Spirulina bioplastic. The ingredients include
lack of bacterial contamination of Spirulina platensis was demonstrated Sorbitol 4.5 g, Gelatin 2.25 g, Glycerol solution 180ml of 1% , Water
by streak plate method using nutrient agar medium. Then Spirulina 240ml. The bioplastic and commercial petrol based plastic was
was transferred to steel containers filled with Zarrouk medium in compared with one another. The plasticizing capacity, moldable
large scale level for analyse the growth ability of Spirulina platensis property of bioplastic and commercial plastic was analyzed. The
and optimize the medium in aim of increase PHB concentration. In ingredient used in preparation of plastic and bioplastic was compared
both level of cultivation frequent mixing is provided, thrice a day for biodegradable property and cost required of production.
minimum. The condition maintained up to the Spirulina float on
above surface of the container (Campbell et al., 1981). Statistical analysis: The correlation between cell dry weight (g
L-1) and PHB production (g L-1) is determined according to
During large-scale level determination amount of PHB is Sperman’s ñ correlation coefficient test (Yavuz Beyalti, 2006).
performed chemically. 10 ml of Spirulina was centrifuged at 6000
rpm for 30 min and pellet was dried for 24 hours at 100ºC. Total (X -Y )2
6∑ 1 1
Spirulina dry weight was determined, the pellet was suspended in ρ = 1 - ___________
sterile water and homogenized then allowed for vortex mixing. n(n2-1)
From 2 ml of suspension 2 ml of 2N HCL was added then heated
for 2 hours in water bath. Then the tube was centrifuged at 6000 Results
rpm for 20 min and 5 ml of chloroform is added and left overnight at The S. platensis was collected and cultured in modified
28ºC on a shaker at 150 rpm. Then centrifuged at 6000 rpm for 20 Zarrouk medium. The growth ability of Spirulina platensis was
min, extracted with 0.1 ml of chloroform and dried at 40ºC. 5 ml of observed as good in both small and large scale level. The dry cell
concentrated sulfuric acid was added to the tube and the mixture weight of Spirulina platensis in 7 days was 0.135 and the PHB
was heated at 100ºC in a water bath for 20 min. After the PHB amount was 0.006. After optimization, in 14 days the amount of cell
crystals were converted to crotonic acid. The PHB content was dry weight and PHB concentration of Spirulina platensis was 0.290
measured at 235 nm in UV spectrophotometer against sulphuric acid and 0.018. The PHB content was increased in high amount. From
blank. From the cell dry weight and PHB content the percentage of the total cell dry weight and PHB content, the percentage of PHB is
PHB produced by the organisms was calculated (Bonartseva, 1987). calculated. In 7 days the percentage of PHB was 5.18% and in 14

Research in Environment and Life Sciences 134 August, 2011

N. Uma Maheswari and K. Ahilandeswari Production of bioplastic

Table-1: Production of PHB in Spirulina platensis whereas Jensen and Sicko in 1966 demonstrated PHB granules in
Sample Cell dry PHB % of PHB the same Cyanobacterium by electron microscopy. Carr was not
(Spirulina platensis) wt. (g L-1) (g L-1) (Cell dry wt.) able to detect any PHB when C. fritschii was grown in the absence
of acetate. Thus, although S.platensis accumulates PHB as does
7 days 0.135 0.006 5.18
C. fritschii, it is different in that reduced carbon compounds need not
14 days 0.290 0.018 6.20
be supplied for PHB accumulation.
Values are represented as Mean
Carr report was slightly differing from Gibson report (1981).
Table - 2: Composition for 1 kg bioplastic and plastic preparation
Because, Gibson reported the PHB could be regarded as a
Composition (g) Bioplastic Plastic fermentation product, and its accumulation in cyanobacterial cells
Sorbitol 225 450 growing photoautotrophically. In mixotrophic mode of growth, the
Gelatin 225 225 presence of acetate in the growth medium is known to increase
S.platensis 225 0 intracellular acetyl-CoA concentration at the expense of the free
Glycerol 1% conc (lr) 18 18 CoA pool and to stimulate the synthesis of lipids. In this study also
Water (lr) 24 24 the Spirulina platensis cultivated in modified Zarrouk medium using
sodium acetate. The intracellular accumulation of PHB induced by
Table - 3: Cost for 1 kg bioplastic and plastic preparation the addition of sodium acetate.
Composition (g) Bioplastic (Rs) Plastic (Rs)
Dawes gave same report in 1986, the synthesis of PHB is
Sorbitol 148 297 a reductive process regulated primarily by 3-ketothiolase. This
Gelatin 405 405 enzyme is powerfully inhibited by high concentration of free
S.platensis 270 0 coenzyme A, but at high concentrations of acetyl- CoA, the inhibitory
Glycerol 1% conc (lr) 80 80
effect of CoA is overcome.
Water (lr) 0 0
Our study was correlated with Emo chiellini et al (2008),
days the PHB was 6.20%. The percentage of PHB produced by the biodegradable thermoplastic composites based on polyvinyl
the organism was considerably high (1.02%) in after treatment alcohol and algae. Fibers of the green alga Ulva armoricana were
(Table-1). The increased concentration was due to increased growth positively evaluated for the production of composites with poly(vinyl
of organism in stationary phase and intracellular accumulation of alcohol) (PVA) as continuous matrix by casting of aqueous
PHB in the presence of sodium acetate. suspensions and compression molding. PVA, Ulva, and starch were
The correlation between cell dry weight and PHB content also successfully processed by the melt in the presence of glycerol.
was calculated as 0.04(ñ). When this value was compared with Positive results were obtained for film-forming properties and
critical value the result was there is no correlation between cell dry mechanical characteristics with limited amount of PVA (40%) attesting
weight and PHB content of the cyanobacteria. for Ulva suitability to be introduced in composites (up to 30%).
Alexander Steinbuchel (1991) reported one of the properties
The bioplastic and plastic was compared for moldable
that distinguish PHB from petroleum-based plastics is their
and plasticizing property. Here the two plastic had good moldable
biodegradability. The PHB are degraded upon subsequent
and plasticizing capacity was observed. Then the bioplastic and
exposure to soil, compost, or marine sediment. Despite their
commercial plastic was compared for biodegradable property and
biodegradability the PHB still have good resistance to water and
production cost required for preparation. The biodegradable
moisture vapor, and are stable under normal storage conditions
property of Spirulina bioplastic was high compare than commercial
and during use.
plastics due to presence of PHB content (6.20%), it was confirmed
by UV spectrophotometry method. The presence of PHB in Spirulina Conclusion
platensis bioplastic increases the biodegradable property.
It is therefore, conclude the biodegradability of bioplastic
By comparing the cost requirement of the bioplastic with was good but the production cost was high due to high cost required
commercial plastic, the commercial plastic was cheaper one because of Spirulina cultivation. Because the Spirulina platensis normally
the synthetic polymers was low price. But S. platensis was high cultivated in high cost chemical medium. However, the
cost, then cultivation of cyanobacteria not a simple process. Cyanobacterium able to grow in cheap organic medium and modified
Therefore, the preparation of bioplastic requires too high cost chemical medium. Zarrouk medium was used for cultivation of
compare to commercial plastics (Table 2 and 3). Spirulina platensis. If cultivate the Spirulina platensis in simple organic
medium in mass cultivation the cost for preparation of Spirulina
Discussion bioplastic could be minimized. If cultivate the cyanobacteria in low
Our study was correlated with Carr et al. (1966) cost organic medium, in future 100% biodegradable bioplastic will
demonstration, the PHB in Chlorogloea fritschii by chemical analysis, be produced by using cyanobacterial starch, cyanobacterial derived

Research in Environment and Life Sciences 135 August, 2011

N. Uma Maheswari and K. Ahilandeswari Production of bioplastic

oil and cyanobacterial PHB. Thus we can use the bioplastics in our Emo Chiellini, Patrizia Cinelli, Vassilka, I. H. and Martina Martera:
Biodegradable Thermoplastic Composites based on polyvinyl alcohol
bulk packaging systems where conventional plastic is basically used
and algae. Biomacromolecules., 9: 1007-1013 (2008).
and save our environment. Falcone, D.B.: Biodegradable plastic in the environment M.Sc., Thesis,
UFSCar, Sao Paulo, Brazil (2004).
It is worth noting that biodegradable plastics cannot yet Gibson, J.: Movement of acetate across the cytoplasmic membrane of the
compete with traditional plastics because of their higher production unicellular cyanobacterium Synechococcus and Aphanocapsa. Arch.
cost. Therefore, the reduction of PHB production costs is presently Microbiol., 130: 175-179 (1981).
Greg Stevens.: HOW TO: make algae bioplastic (http://green-plastic.net)
one of the major topics in biopolymer research. (2010).
Jensen, T. E. and Sicko. L. M.: Fine structure of poly-â-hydroxy butyric
References acid in a blue-green alga, Chlorogloea fritschii. J. Bacteriol., 106:
Alexander Steinbuchel.: Polyhydroxy alkonates In: M. David Byron (Ed.) 683-686 (1971).
Biomaterials. Stockton Press, New York (1991). Lemoigne, M.: Produits de deshydration et de polymerization de lactic â-
Bonartseva, G. A. and Myshkina, V. L.: Fluorescence intensity of strain of oxy butyrique. Bull. Soc. Chim. Boil., 8: 770-782 (1926).
nodule bacteria(Rhizobium meliloti and Rhizobium phaseoli) differing Lemoigne, M.: Polyhydroxyalkanoates: Characteristics and Processing.
in activity, grown in the presence of the lipophilic vital stain phosphine. Ann Inst Pasteur., 41:148 (1927).
Microbiology., 54: 535-541 (1985). Massimo Vincenzini, Claudio Sili, Roberto DePhillippis, Alba Ena and
Campbell, J., Stevensm, S. E. and Balkwill, D. L.: Accumulation of poly Richardo Materassi.: Occurance of Poly-α-Hydroxybutyrate in
hydroxybutyrate in spirulina platensis. J. Bacteriol., 149: 361-363 (1982). Spirulina species. J. Bacteriol., 172: 2791-2792 (1990).
Carr, N. G.: The occurrence of poly â hydroxybutyrate in the blue-green- Okada, M.: Chemical synthesis of biodegradable polymers. Progress in
alga, Chlorogloea fritschii. Biochim. Biophys., 120: 308-310 (1966). Polymer Science, 27: 87-133 (2002).
Costa, J. A., Colla, L. M. and Duarte Filho, P. F.: Improving Spirulina Pandey, J. P., Amit Tiwari. And Mishra, M.: Evaluation of biomass production
Platensis Biomass yeild using a fed-batch process. Bioprocess of Spirulina maxima. J. Algal Biomass utln., 1: 70-81 (2010).
Technol., 92: 237-241 (2004). Yavuz Beyalti, Belma Aslim, Hikmet Katircioglu and Zehra Nur Yuksekdag.:
Dawes, E.A.: Microbial energetics, Blackie and Son Ltd., Glassgow, Determination of PHB Production in Different Growth Medium by
Scotland (1986). Cyanobacteria. Pak. J. Biol. Sci., 9: 45-47 (2006).

Research in Environment and Life Sciences 136 August, 2011